Fig. 4: Multiplexed detection and spatial identification using the SFB system.
From: Spatial fluorescence barcode by transiently luminescent DNA beads
a Generation of the mSFBC. The sequential introduction of Targets I-VII enables cyclic fluorescence imaging. TMSD reactions between targets and their corresponding Y-shaped probes on polystyrene beads transiently activate fluorescence, revealing the spatial positions of target-associated DNA beads. ExoIII-mediated target degradation resets the fluorescence signal, completing each cycle. Each type of DNA target (TLDBs) assigns a distinct pseudo-color. Merged fluorescence images of all SFBs corresponding to TLDBs generate the mSFBC reference, encoding spatial information for each target. b TLDB fluorescence images of an unknown sample SFB compared to the mSFBC, identifying Targets I, II, III, and VI. c Bead-by-bead comparison of TLDB fluorescence images in the unknown sample SFB (left) and the corresponding mSFBC (right). Data are mean ± S.D. (n = 3 independent experiments). The figure contains elements created by BioRender. d Colocalization analysis across three independent imaging fields. Each dot represents a PCC value between corresponding DNA beads in the unknown sample SFB and mSFBC reference. mPCC values for DNA beads I-VII are shown above each group. Error bars represent standard deviation. Scale bar: 5 μm. Source data can be found in the Source Data file.
